FIGURE 4.

Increasing Ski protein levels under HG conditions increased the binding of Ski to FoxO1 and thus inhibited acetylation of FoxO1. Fibroblasts were infected with Ad‐Ski or Ad‐EGFP for 48 hours, combined with HG treatment for 48 hours. A, IP with an anti‐FoxO1 antibody followed by immunoblotting with anti‐acetyl‐lysine (ac‐lysine) or total FoxO1 antibodies. B,C, IP of Ski and FoxO1 using an anti‐Ski or anti‐FoxO1 antibody as the IP antibody and Western blotting with an anti‐FoxO1 or anti‐Ski antibody. Rabbit IgG served as a negative control. Input is the total cell lysate of the control without anti‐FoxO1 or anti‐Ski immunoprecipitation. The data are presented as the mean ± SEM, and each experiment was repeated three times. *P < .05, **P < .01, ***P < .001 compared to the DMEM + NG group; ^#P < .05, ^##P < .01, ^###P < .001 compared to the DMEM + HG group, one‐way ANOVA, Dunnett's post hoc tests